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  • Title: The transition to an elongation complex by T7 RNA polymerase is a multistep process.
    Author: Bandwar RP, Ma N, Emanuel SA, Anikin M, Vassylyev DG, Patel SS, McAllister WT.
    Journal: J Biol Chem; 2007 Aug 03; 282(31):22879-86. PubMed ID: 17548349.
    Abstract:
    During the transition from an initiation complex to an elongation complex (EC), T7 RNA polymerase undergoes major conformational changes that involve reorientation of a "core" subdomain as a rigid body and extensive refolding of other elements in the 266 residue N-terminal domain. The pathway and timing of these events is poorly understood. To examine this, we introduced proline residues into regions of the N-terminal domain that become alpha-helical during the reorganization and changed the charge of a key residue that interacts with the RNA:DNA hybrid 5 bp upstream of the active site in the EC but not in the initiation complex. These alterations resulted in a diminished ability to make products >5-7 nt and/or a slow transition through this point. The results indicate that the transition to an EC is a multistep process and that the movement of the core subdomain and reorganization of certain elements in the N-terminal domain commence prior to promoter release (at 8-9 nt).
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